WU多瘤病毒和Saffold心病毒在儿童患者中的流行规律与基因特征研究
发布时间：2018-09-06 16:26
【摘要】：研究背景新发传染病始终威胁着人类的健康,发现和鉴定新病毒以及确定新病毒与疾病的关系是预防、诊断和治疗新发病毒性传染病的首要任务。近十多年来,随着生物技术的发展,越来越多的病毒被发现,如冠状病毒NL63、HKU1、MERS病毒、埃博拉病毒、博卡病毒和偏肺病毒等。尽管近年来对疾病病毒感染的病原学已有较深入的研究,但目前仍有一部分的疾病病因不明,部分未知的疾病病因可能是由未被发现的病毒所引起。WU多瘤病毒是2007年5月美国华盛顿大学医学院科学家Gaynor通过高通量序列测定,从一名患有肺炎的3岁儿童的鼻咽抽吸物标本中发现的一种新病毒。Saffold心病毒于1981年从一个8个月大的不明原因发热病人的粪便中首次分离。在2007年,通过非序列依赖的单引物扩增,确定为心病毒属中感染人类的新种。研究报道,在患者的鼻咽抽吸物、粪便、血清等标本中可以检测出WU多瘤病毒和Saffold心病毒,因此可能是引起疾病的病原体之一。由于WU多瘤病毒和Saffold心病毒是新发现的病毒,近年来越来越引起广泛的关注。当前国内对于这两种病毒在中国地区的流行规律,致病性和基因特征缺乏了解,大多数的研究是仅仅局限在一种疾病中检测分析这两种病毒,缺乏对于病毒的流行规律、临床特征和基因特征进行分析。目的本研究通过对重庆医科大学附属儿童医院的呼吸道感染患者鼻咽抽吸物标本、急性腹泻患者和手足口病患者的粪便标本进行病原体检测,对新发现的WU多瘤病毒、Saffold心病毒在儿童患者中的流行规律和基因进化特征进行系统性的分析,从而为下一步的病毒感染治疗、疫情控制提供科学依据和参考意见。方法以重庆医科大学附属儿童医院作为哨点监测医院,收集2012年1月到2015年12月的呼吸道感染患者的鼻咽抽吸物标本、急性腹泻患者的粪便标本和手足口病患者的粪便标本。通过采用PCR,RT-PCR,Real-Time PCR,Real-Time RT-PCR等方法检测不同标本中的病毒。呼吸道感染患者鼻咽抽吸物标本检测的常规病毒包括人腺病毒(human adenovirus,HAd V)、流感病毒(influenza virus,Flu)、人鼻病毒(human rhinovirus,HRV/HEV)、呼吸道合胞病毒(respiratory syncytial virus,RSV)、偏肺病毒(metapneumovirus,MPV)、副流感病毒(parainfluenza,PIV)、人博卡病毒(human bocavirus,HBo V)和人冠状病毒(human coronavirus,HCo V)。急性腹泻患者粪便标本检测的常规病毒包括轮状病毒(rotavirus,Rt V)、诺如病毒(norovirus,No V)、腺病毒(adenovirus,Ad V)、扎如病毒(sapovirus,Sp V)和星状病毒(astrovirus,At V)。手足口病患者粪便标本中检测的常规病毒是肠道病毒(Enterovirus,EV)、肠道病毒71型(Enterovirus 71,EV71)和柯萨奇病毒A16型(Coxsackievirus A16,CVA16)。呼吸道感染患者鼻咽抽吸物标本、急性腹泻患者粪便标本和手足口病患者粪便标本均检测WU多瘤病毒(WU Polyomavirus,WUPy V)和Saffold心病毒(Saffold Cardiovirus,SAFV)。对WU多瘤病毒阳性的标本进行全基因组扩增,Saffold心病毒阳性的样本进行VP1片段扩增并分型,并进行系统发育分析。使用Epidata3.1录入数据,采用SPSS 20.0软件进行统计分析,检验水准α=0.05。构建系统发育进化树采用软件Mega 7.0,使用maximum likelihood法,bootstrap值设定为1000进行构建和检验。选择压力分析采用FEL、IFEL、MEME、SLAC四种方法筛选,同一位点至少在三种方法中检出则判定为正选择位点,使用Data Monkey网站提交序列进行分析(http://www.datamonkey.org/)。结果1.WU多瘤病毒在儿童患者中的流行规律与基因特征研究(1)本研究共检测出WUPy V阳性170例,总检出率为4.7%。其中呼吸道感染患者鼻咽抽吸物标本中WUPy V阳性127例(7.8%),急性腹泻患者粪便标本中WUPy V阳性25例(2.4%),手足口病患者的粪便标本中WUPy V阳性18例(1.9%),三者的阳性检出率之间差异具有统计学意义(P0.001),呼吸道感染患者WUPy V阳性率最高。(2)在三种类型的病例中,WUPy V复合感染率为79.4%(135/170),合并一种病毒感染97例,合并两种病毒感染31例,合并三种病毒及以上感染7例,单独感染WUPy V的有35例。呼吸道感染患者复合感染率为79.5%(101/127),复合感染发生最多的病毒是HRV/HEV(28例);急性腹泻患者复合感染率为76.0%(19/25),复合感染发生最多的病毒是Rt V(11例);手足口病患者复合感染率为83.4%(15/18),复合感染发生最多的病毒是EV71(9例)。(3)从检测阳性率的时间分布看,呼吸道感染患者在2013年5-6月WUPy V阳性检出率最高,急性腹泻患者和手足口病患者在2013年7月出现一次检测阳性率高峰,提示在这一时间段,可能出现WUPy V感染的暴发。(4)从临床症状上看,在呼吸道感染患者中,WUPy V复合感染的患者出现咳嗽,干Up音,湿Up音等症状的频率大于WUPy V单独感染的患者(P0.05)。在上呼吸道感染患者中WUPy V感染率为11.4%,且和非上呼吸道感染患者相比,差异具有统计学意义(P=0.018,OR=1.770,95%CI:1.103-2.842),肺炎患者中WUPy V感染率为7.8%。(5)本研究成功扩增57株WUPy V全基因组序列,同源性在98.7%-100%。同Gen Bank上已经上传的其他地区的序列做系统进化分析,显示形成了I、II、III三个分支。本次研究中的序列聚集在Ia、Ic和IIIc,并且形成了一个新的亚支IIIc。来自呼吸道感染患者的35株序列,分别聚集于Ia(20株),Ic(8株),IIIc(7株)。来自急性腹泻患者标本中的12株序列,全部分布在Ia分支。来自手足口病患者标本的10株序列,8株在Ia,2株在IIIc。(6)WUPy V选择压力分析显示VP1片段的第82号位点为正选择位点,VP2,VP3,STAg,LTAg均未发现正选择位点。2.Saffold心病毒在儿童患者中的流行规律与基因特征研究(1)本研究中共检测出SAFV阳性190例,总检出率为2.0%。其中在呼吸道感染患者鼻咽抽吸物标本中检测出44例(1.3%),急性腹泻患者粪便标本中检测出28例(0.9%),手足口病患者的粪便中检测出118例(3.5%),三者的阳性检出率不同,差异具有统计学意义(P0.001),手足口病患者SAFV阳性率最高。(2)SAFV总的复合感染率为73.2%(139/190),合并一种病毒感染116例,合并两种病毒感染18例,合并三种病毒及以上感染5例,SAFV单独感染51例。呼吸道感染患者复合感染率为84.1%(37/44),复合感染发生最多的病毒是RSV(12例);急性腹泻患者复合感染率为67.9%(19/28),复合感染发生最多的病毒是Rt V(16例);手足口病患者复合感染率为70.3%(83/118),复合感染发生最多的病毒是EV71(36例)。(3)呼吸道感染患者中,大于36个月年龄组患者的SAFV阳性率高于1-6个月年龄组(0.95%vs 2.9%,P=0.005,OR=3.047,95%CI:1.396-6.651)。上呼吸感染患者中SAFV的阳性率为1.3%,肺炎患者中SAFV的阳性率为1.1%,且和非肺炎患者相比,差异具有统计学意义(P=0.001,OR=0.308,95%CI:0.152-0.627)。(4)手足口病患者中,具有神经系统症状的患者SAFV的阳性率高于无神经系统症状的患者(P=0.040,OR=1.475,95%CI:1.016-2.140),重度手足口病患者的阳性率高于轻度手足口患者(P=0.021,OR=1.535,95%CI:1.063-2.219)。(5)EV71和SAFV复合感染与EV71单独感染的患者相比较,临床重症的发生率差异具有统计学意义,EV71和SAFV复合感染更容易加重病情(P=0.007)。(6)本研究中通过对SAFV阳性标本进行VP1片段扩增,获得151株VP1基因序列,系统进化分析显示4种型别的存在:SAFV-1型(17株),SAFV-2型(70株),SAFV-3型(59株),SAFV-6型(5株)。对各种型别分别进行系统进化分析,显示序列具有一定的聚集性和地理分布特征。结论1.WU多瘤病毒在儿童患者中的流行规律与基因特征研究(1)首次在手足口病患者中检测到WUPy V,其阳性率为1.9%。2013年5-7月三种症候群患者中WUPy V阳性率均较高,提示在这一时间段WUPy V可能出现暴发。(2)获得了中国地区57株WUPy V全基因组序列。通过系统发育分析表明,本研究中的序列形成了一个新的亚支IIIc,序列分布具有地域特征,同时可能存在地域传播。手足口病患者粪便标本中的10株全基因组序列,其主要型别是Ia型和IIIc型。急性腹泻患者粪便标本中的12株全基因组序列全部为Ia型。(3)WUPy V基因突变不仅存在纯化选择作用,也有正向选择作用的影响。2.Saffold心病毒在儿童患者中的流行规律与基因特征研究(1)首次在手足口患者中检测到SAFV,其阳性率为3.5%,高于呼吸道感染患者和急性腹泻患者。(2)在手足口病患者中,SAFV可能和神经系统症状、重度手足口病相关,同时EV71和SAFV复合感染可能会加重手足口病患者的病情。(3)系统进化分析显示,本研究中的SAFV序列分别为SAFV-1型,SAFV-2型,SAFV-3型,SAFV-6型,不同的基因型均具有一定的聚集性和地理分布特征。
[Abstract]:Background Emerging infectious diseases have always threatened human health. The discovery and identification of new viruses and the determination of the relationship between new viruses and diseases are the primary tasks in the prevention, diagnosis and treatment of new viral infectious diseases. Ebola virus, Boca virus and hemipneumonia virus. Although the etiology of disease virus infection has been studied in depth in recent years, there are still some unknown causes of disease, some unknown causes of disease may be caused by undetected viruses. WU polyoma virus is May 2007, the United States University of Washington Medical School Department. Saffold's heart virus was first isolated in 1981 from the stool of an eight-month-old fever patient of unknown origin. In 2007, it was identified as a cardiovirus by non-sequence-dependent single-primer amplification. It is reported that WU polyomavirus and Saaffold heart virus can be detected in nasopharyngeal aspirates, feces and serum of patients, so they may be one of the pathogens causing the disease. As WU polyomavirus and Saaffold heart virus are newly discovered viruses, they have attracted more and more attention in recent years. The epidemic regularity, pathogenicity and genetic characteristics of the two viruses in China are poorly understood. Most of the studies are limited to one disease. The epidemic regularity, clinical characteristics and genetic characteristics of the two viruses are not analyzed. Nasopharyngeal aspirate specimens from patients with respiratory tract infection, fecal specimens from patients with acute diarrhea and hand-foot-mouth disease were tested for pathogens, and the epidemic regularity and gene evolution characteristics of newly discovered WU polyoma virus and Saffold heart virus in children were systematically analyzed, so as to provide further treatment for viral infection and control of the epidemic situation. Methods Nasopharyngeal aspirates, feces of patients with acute diarrhea and feces of patients with hand-foot-mouth disease were collected from the children's Hospital Affiliated to Chongqing Medical University from January 2012 to December 2015. PCR, RT-PCR, Real-Time PCR, Real-Ti were used. The routine viruses detected in nasopharyngeal aspirates of patients with respiratory tract infection include human adenovirus (HAd V), influenza virus (Flu), human rhinovirus (HRV / HEV), respiratory syncytial virus (RSV), hemipneumonia virus (met). APneumovirus (MPV), parainfluenza virus (PIV), human bocavirus (HBo V) and human coronavirus (HCo V). Conventional viruses detected in stool samples from patients with acute diarrhea include rotavirus (Rt V), norovirus (No), adenovirus (Ad V), sapoviruses (Sapoviruses). Enterovirus (EV), Enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) are the common viruses detected in stool specimens from patients with HFMD. Nasopharyngeal aspirate specimens from patients with respiratory tract infections, stool specimens from patients with acute diarrhea, and hand, foot and mouth specimens from patients with HFMD WU polyomavirus (WUPy V) and Saffold Cardiovirus (SAFV) were detected in fecal specimens of the patients. WU polyomavirus positive specimens were amplified by whole genome amplification, Saffold cardiovirus positive specimens were amplified by VP1 fragment amplification, typing and phylogenetic analysis. Epidata 3.1 data were recorded and SPS was used S 20.0 software for statistical analysis, test level alpha = 0.05. Construction of phylogenetic tree using software Mega 7.0, using maximum likelihood hood method, bootstrap value set to 1000 for construction and testing. Selective pressure analysis using FEL, IFEL, MEME, SLAC four methods screening, at least three methods of the same site detection is determined to be a positive choice. Results 1. Epidemiological and genetic characteristics of WU polyoma virus in children (1) This study detected 170 cases of WUPy V positive, the total detection rate was 4.7%. Of them, 127 cases (7.8%) were WUPy V positive in nasopharyngeal aspirates of respiratory tract infection patients. There were 25 (2.4%) WUPy V positive stool specimens from patients with acute diarrhea and 18 (1.9%) WUPy V positive stool specimens from patients with hand-foot-mouth disease. The positive rates of WUPy V were statistically significant (P 0.001). The positive rate of WUPy V in patients with respiratory tract infection was the highest. (2) The combined infection rate of WUPy V was 79.4% (135/170) among the three types of cases. There were 97 cases with one virus infection, 31 cases with two viruses infection, 7 cases with three or more viruses infection and 35 cases with WUPy V infection alone. The most common viruses were Rt V (11 cases), hand-foot-mouth disease (83.4% (15/18) and EV71 (9 cases). (3) According to the time distribution of the positive rate, the positive rate of WUPy V was the highest in patients with respiratory tract infection from May to June 2013, and acute diarrhea and hand-foot-mouth disease (HFMD) had a test in July 2013. The positive rate of WUPy V infection was higher than that of WUPy V infection alone (P 0.05). The infection rate of WUPy V was 11.4% in patients with upper respiratory tract infection. The WUPy V infection rate was 7.8%. (5) 57 WUPy V genome sequences were successfully amplified with homology of 98.7%-100%. Phylogenetic analysis showed that the WUPy V infection rate was significantly different from that of non-upper respiratory tract infection patients (P = 0.018, OR = 1.770, 95% CI: 1.103-2.842). In this study, the sequences were clustered in Ia, Ic and IIC, and a new subbranch IIIc was formed. 35 strains from patients with respiratory tract infection were clustered in Ia (20 strains), Ic (8 strains), and IIC (7 strains). 12 strains from patients with acute diarrhea were all distributed in Ia. Sequences of 10 strains, 8 strains in Ia, 2 strains in IIIc. (6) WUPy V selection pressure analysis showed that VP1 fragment No. 82 was a positive selection site, VP2, VP3, STAg, LTAg were not found positive selection sites. Among them, 44 cases (1.3%) were detected in nasopharyngeal aspirates of patients with respiratory tract infection, 28 cases (0.9%) in feces of patients with acute diarrhea and 118 cases (3.5%) in feces of patients with hand-foot-mouth disease. The positive rates of the three cases were different, and the difference was statistically significant (P 0.001). The combined infection rate was 73.2% (139/190), 116 cases complicated with one virus infection, 18 cases complicated with two viruses infection, 5 cases complicated with three viruses and more, 51 cases infected with SAFV alone. (3) In patients with respiratory tract infection, the positive rate of SAFV in patients older than 36 months was higher than that in 1-6 months (0.95% vs 2.9%, P = 0.005, OR = 3.047, 95% CI: 1.396-6.651). The positive rate of SAFV was 1.3% in patients with upper respiratory infection and 1.1% in patients with pneumonia, and the difference was statistically significant (P = 0.001, OR = 0.308, 95% CI: 0.152-0.627). (4) In patients with HFMD, the positive rate of SAFV in patients with neurological symptoms was higher than that in patients without neurological symptoms (P = 0.040, O = 0, O = 0.040). R = 1.475, 95% CI: 1.016-2.140, and the positive rate of severe HFMD patients was higher than that of mild HFMD patients (P = 0.021, OR = 1.535, 95% CI: 1.063-2.219). (5) Comparing with EV71 and SAFV infection alone, the incidence of severe clinical infections was significantly different. EV71 and SAFV infection were more likely to aggravate the disease (P = 0.007). In this study, 151 VP1 gene sequences were obtained from SAFV-positive specimens. Phylogenetic analysis showed that there were four types: SAFV-1 (17 strains), SAFV-2 (70 strains), SAFV-3 (59 strains) and SAFV-6 (5 strains). Phylogenetic analysis showed that the sequences were clustered and geographically distributed. The positive rate of WUPy V was 1.9%. The positive rate of WUPy V was higher among the three groups of patients from May to July 2013, suggesting that there might be an outbreak of WUPy V in this period. (2) 57 strains of WUPy V were obtained in China. Genomic sequence. Phylogenetic analysis showed that the sequence in this study formed a new subbranch IIIc with regional characteristics and possible regional transmission. 10 complete genomic sequences of fecal samples from patients with hand-foot-mouth disease were of type Ia and type IIIc. 12 of the fecal samples from patients with acute diarrhea were of type Ia and type IIIc. The whole genome sequence was typeIa. (3) WUPy V gene mutation not only had purification selection effect, but also had positive selection effect. (2) Saffold heart virus epidemiology and genetic characteristics in children (1) SAFV was detected in hand, foot and mouth patients for the first time, the positive rate was 3.5%, higher than that in respiratory tract infection patients and acute diarrhea patients. (2) In HFMD patients, SAFV may be associated with neurological symptoms, severe HFMD, and the combined infection of EV71 and SAFV may aggravate the condition of HFMD patients. (3) Phylogenetic analysis showed that the SAFV sequences in this study were SAFV-1, SAFV-2, SAFV-3, SAFV-6, and different genotypes all had certain genotypes. The characteristics of clustering and geographical distribution.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R725.1;R181.3
本文编号：2226903
[Abstract]:Background Emerging infectious diseases have always threatened human health. The discovery and identification of new viruses and the determination of the relationship between new viruses and diseases are the primary tasks in the prevention, diagnosis and treatment of new viral infectious diseases. Ebola virus, Boca virus and hemipneumonia virus. Although the etiology of disease virus infection has been studied in depth in recent years, there are still some unknown causes of disease, some unknown causes of disease may be caused by undetected viruses. WU polyoma virus is May 2007, the United States University of Washington Medical School Department. Saffold's heart virus was first isolated in 1981 from the stool of an eight-month-old fever patient of unknown origin. In 2007, it was identified as a cardiovirus by non-sequence-dependent single-primer amplification. It is reported that WU polyomavirus and Saaffold heart virus can be detected in nasopharyngeal aspirates, feces and serum of patients, so they may be one of the pathogens causing the disease. As WU polyomavirus and Saaffold heart virus are newly discovered viruses, they have attracted more and more attention in recent years. The epidemic regularity, pathogenicity and genetic characteristics of the two viruses in China are poorly understood. Most of the studies are limited to one disease. The epidemic regularity, clinical characteristics and genetic characteristics of the two viruses are not analyzed. Nasopharyngeal aspirate specimens from patients with respiratory tract infection, fecal specimens from patients with acute diarrhea and hand-foot-mouth disease were tested for pathogens, and the epidemic regularity and gene evolution characteristics of newly discovered WU polyoma virus and Saffold heart virus in children were systematically analyzed, so as to provide further treatment for viral infection and control of the epidemic situation. Methods Nasopharyngeal aspirates, feces of patients with acute diarrhea and feces of patients with hand-foot-mouth disease were collected from the children's Hospital Affiliated to Chongqing Medical University from January 2012 to December 2015. PCR, RT-PCR, Real-Time PCR, Real-Ti were used. The routine viruses detected in nasopharyngeal aspirates of patients with respiratory tract infection include human adenovirus (HAd V), influenza virus (Flu), human rhinovirus (HRV / HEV), respiratory syncytial virus (RSV), hemipneumonia virus (met). APneumovirus (MPV), parainfluenza virus (PIV), human bocavirus (HBo V) and human coronavirus (HCo V). Conventional viruses detected in stool samples from patients with acute diarrhea include rotavirus (Rt V), norovirus (No), adenovirus (Ad V), sapoviruses (Sapoviruses). Enterovirus (EV), Enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) are the common viruses detected in stool specimens from patients with HFMD. Nasopharyngeal aspirate specimens from patients with respiratory tract infections, stool specimens from patients with acute diarrhea, and hand, foot and mouth specimens from patients with HFMD WU polyomavirus (WUPy V) and Saffold Cardiovirus (SAFV) were detected in fecal specimens of the patients. WU polyomavirus positive specimens were amplified by whole genome amplification, Saffold cardiovirus positive specimens were amplified by VP1 fragment amplification, typing and phylogenetic analysis. Epidata 3.1 data were recorded and SPS was used S 20.0 software for statistical analysis, test level alpha = 0.05. Construction of phylogenetic tree using software Mega 7.0, using maximum likelihood hood method, bootstrap value set to 1000 for construction and testing. Selective pressure analysis using FEL, IFEL, MEME, SLAC four methods screening, at least three methods of the same site detection is determined to be a positive choice. Results 1. Epidemiological and genetic characteristics of WU polyoma virus in children (1) This study detected 170 cases of WUPy V positive, the total detection rate was 4.7%. Of them, 127 cases (7.8%) were WUPy V positive in nasopharyngeal aspirates of respiratory tract infection patients. There were 25 (2.4%) WUPy V positive stool specimens from patients with acute diarrhea and 18 (1.9%) WUPy V positive stool specimens from patients with hand-foot-mouth disease. The positive rates of WUPy V were statistically significant (P 0.001). The positive rate of WUPy V in patients with respiratory tract infection was the highest. (2) The combined infection rate of WUPy V was 79.4% (135/170) among the three types of cases. There were 97 cases with one virus infection, 31 cases with two viruses infection, 7 cases with three or more viruses infection and 35 cases with WUPy V infection alone. The most common viruses were Rt V (11 cases), hand-foot-mouth disease (83.4% (15/18) and EV71 (9 cases). (3) According to the time distribution of the positive rate, the positive rate of WUPy V was the highest in patients with respiratory tract infection from May to June 2013, and acute diarrhea and hand-foot-mouth disease (HFMD) had a test in July 2013. The positive rate of WUPy V infection was higher than that of WUPy V infection alone (P 0.05). The infection rate of WUPy V was 11.4% in patients with upper respiratory tract infection. The WUPy V infection rate was 7.8%. (5) 57 WUPy V genome sequences were successfully amplified with homology of 98.7%-100%. Phylogenetic analysis showed that the WUPy V infection rate was significantly different from that of non-upper respiratory tract infection patients (P = 0.018, OR = 1.770, 95% CI: 1.103-2.842). In this study, the sequences were clustered in Ia, Ic and IIC, and a new subbranch IIIc was formed. 35 strains from patients with respiratory tract infection were clustered in Ia (20 strains), Ic (8 strains), and IIC (7 strains). 12 strains from patients with acute diarrhea were all distributed in Ia. Sequences of 10 strains, 8 strains in Ia, 2 strains in IIIc. (6) WUPy V selection pressure analysis showed that VP1 fragment No. 82 was a positive selection site, VP2, VP3, STAg, LTAg were not found positive selection sites. Among them, 44 cases (1.3%) were detected in nasopharyngeal aspirates of patients with respiratory tract infection, 28 cases (0.9%) in feces of patients with acute diarrhea and 118 cases (3.5%) in feces of patients with hand-foot-mouth disease. The positive rates of the three cases were different, and the difference was statistically significant (P 0.001). The combined infection rate was 73.2% (139/190), 116 cases complicated with one virus infection, 18 cases complicated with two viruses infection, 5 cases complicated with three viruses and more, 51 cases infected with SAFV alone. (3) In patients with respiratory tract infection, the positive rate of SAFV in patients older than 36 months was higher than that in 1-6 months (0.95% vs 2.9%, P = 0.005, OR = 3.047, 95% CI: 1.396-6.651). The positive rate of SAFV was 1.3% in patients with upper respiratory infection and 1.1% in patients with pneumonia, and the difference was statistically significant (P = 0.001, OR = 0.308, 95% CI: 0.152-0.627). (4) In patients with HFMD, the positive rate of SAFV in patients with neurological symptoms was higher than that in patients without neurological symptoms (P = 0.040, O = 0, O = 0.040). R = 1.475, 95% CI: 1.016-2.140, and the positive rate of severe HFMD patients was higher than that of mild HFMD patients (P = 0.021, OR = 1.535, 95% CI: 1.063-2.219). (5) Comparing with EV71 and SAFV infection alone, the incidence of severe clinical infections was significantly different. EV71 and SAFV infection were more likely to aggravate the disease (P = 0.007). In this study, 151 VP1 gene sequences were obtained from SAFV-positive specimens. Phylogenetic analysis showed that there were four types: SAFV-1 (17 strains), SAFV-2 (70 strains), SAFV-3 (59 strains) and SAFV-6 (5 strains). Phylogenetic analysis showed that the sequences were clustered and geographically distributed. The positive rate of WUPy V was 1.9%. The positive rate of WUPy V was higher among the three groups of patients from May to July 2013, suggesting that there might be an outbreak of WUPy V in this period. (2) 57 strains of WUPy V were obtained in China. Genomic sequence. Phylogenetic analysis showed that the sequence in this study formed a new subbranch IIIc with regional characteristics and possible regional transmission. 10 complete genomic sequences of fecal samples from patients with hand-foot-mouth disease were of type Ia and type IIIc. 12 of the fecal samples from patients with acute diarrhea were of type Ia and type IIIc. The whole genome sequence was typeIa. (3) WUPy V gene mutation not only had purification selection effect, but also had positive selection effect. (2) Saffold heart virus epidemiology and genetic characteristics in children (1) SAFV was detected in hand, foot and mouth patients for the first time, the positive rate was 3.5%, higher than that in respiratory tract infection patients and acute diarrhea patients. (2) In HFMD patients, SAFV may be associated with neurological symptoms, severe HFMD, and the combined infection of EV71 and SAFV may aggravate the condition of HFMD patients. (3) Phylogenetic analysis showed that the SAFV sequences in this study were SAFV-1, SAFV-2, SAFV-3, SAFV-6, and different genotypes all had certain genotypes. The characteristics of clustering and geographical distribution.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R725.1;R181.3
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